A modular genetic code expansion approach to site-specific lysine acylations
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Abstract
ABSTRACT Lysine acylations are a diverse class of post-translational modifications (PTMs) that dynamically regulate protein function. Access to homogenous, site-specifically modified proteins is essential for dissecting their molecular roles, yet remains challenging with traditional methods. Here we present a modular strategy that combines genetic code expansion (GCE) with a chemoselective amide bond-forming reaction to install a wide range of lysine acylations directly onto folded proteins. Key to this approach is the genetic encoding of N ε -methoxylysine, which reacts efficiently with N -methyliminodiacetyl (MIDA) acylboronates to generate diverse acyl lysine modifications from a single protein precursor, including several acyl PTMs not previously accessible through GCE. This mild and broadly applicable methodology enables systematic studies of lysine acylation on multiple target proteins. We demonstrate its utility by probing the effects of defined acyl modifications on enzymatic activity, protein-RNA interactions and deacylase-mediated regulation. Together, this platform establishes a versatile strategy to interrogate the functional consequences of this wide-spread class of PTMs.
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Cites
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- Authors
- Knecht, P., Nguyen, T.-A., & Lang, K.
- Affiliations
- ETH Zurich (CH)
- Corresponding author
- Lang, Kathrin
- Venue
- Chem
- Publisher
- Elsevier BV
- Published
- 2026-05
- Volume
- 12
- Issue
- 5
- Pages
- 102919
- ISSN
- 2451-9294 (ISSN-L)
- OpenAlex
- W7133318153
- Cited by
- 2
- References
- 6
- Citations by year
- 2026: 2
- FWCI
- 3.50
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- Chem (publishedVersion, cc-by)
- bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion)
- Repository for Publications and Research Data (ETH Zurich) (publishedVersion, cc-by)
- ETH Zürich Research Collection (cc-by)
- Funders
- H2020 European Research Council (101003289)
- Horizon 2020 Framework Programme
- European Research Council
- Eidgenössische Technische Hochschule Zürich
- Topics
- Physical Sciences › Chemistry › Organic Chemistry › Click Chemistry and Applications
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA and protein synthesis mechanisms
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Chemical Synthesis and Analysis
- Keywords
- lysine acylation, genetic code expansion, site-specific protein modification, post-translational modification, RNA-protein interactions, enzyme activity
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